LOCUS       sequence-479832-        5654 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express TREM2-LgBiT Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: TREM2-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1534..1552)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1535..1552)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1672..1975
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2130..2150
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2183..2230
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2262..3056
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2316..2335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2926..2945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3273..3818
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3719..3738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3935..3952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4102..4150
                     /note="synthetic polyadenylation signal"
     CDS             complement(4174..5211)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4187..4195
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(5242..5599)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5256..5391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5408..5599
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5556..5576
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaaccgtgt ctctcagccc tggcagagtt tggagctgat
      841 accctgggtc atggccacag tccagttcac tgggtggatg tgtccctggc ttctgtccat
      901 gccaggctgc agcccagagg gcgctggctg ctagaatctt gatgagaaag atgcaggcca
      961 ggaggagaag gatggaagtg ggtgggaagg ggatttctcc ttccaagagg ctcctggaga
     1021 tgctgtgctc cacatgggca tcctcgaagc tctcagactc cccggggaac cagagatctc
     1081 cagcatcccg gtgatccagg gggtctgcca gcacctccac caggaccttc ctgagggtgt
     1141 cagcctcact gccatggagg ctctggcact ggtagagacc cgcatcatgg ggttgtagat
     1201 tccgcagcgt aatggtgaga gtgccaccca gggtatcgtc tgtgatggct gtgctcccat
     1261 tccacctcct caggaaggac agcagccaca agttgtgcgt gctgaccaca cgctggcatg
     1321 ggcccttctc tcccagctgg cggcaccagg ccttgcgcct cccccagtgc ttcatggagt
     1381 cataggggca agacacctgc agggactggc ccgccacgcc ctggaacact gtggtgttgt
     1441 gggctccgga cagctctgtg acaaagagta agatgagcag ccggagaggc tccatggcga
     1501 tcgctagctg caaaaagaac aagtagcttg tattctatag tgtcacctaa atgggcgatc
     1561 tgacggttca ctaaaccagc tctgcttata taggcctccc tggcgcgcca ctagttatta
     1621 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1681 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1741 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1801 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     1861 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1921 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1981 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     2041 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     2101 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     2161 ggtcaaatcg aattctaata ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     2221 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     2281 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2341 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2401 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     2461 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2521 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2581 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2641 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     2701 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     2761 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     2821 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     2881 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     2941 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3001 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     3061 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgtca aacatgagaa ttacaactta
     3181 tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag
     3241 aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc
     3301 tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct
     3361 accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt
     3421 tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt
     3481 ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc
     3541 ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg
     3601 aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg
     3661 gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg
     3721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga
     3781 tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct
     3841 cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc
     3901 catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga
     3961 atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca
     4021 tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct
     4081 agcgctgatg tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     4141 atattttatt gcggccatcg tgatggctag agtctaaact cgacctacct ccttagcgcg
     4201 cggtcgtgta ctgggccggc ggttgccgct gtcggccagc acctcgacgc agccgtcggt
     4261 ccatacggct gcgctccggc gagcgatttg tgtgcgaccg acggtgccgg ccccgctgcg
     4321 gacgatggca tcgcagcggc cttgagccca ggcagcatcg tcgaagttgc cgtccacgag
     4381 gctctggtag agctgatcca ggccgatgcg cagcatgtag gctcgcagac gagggctgcc
     4441 ggccagctcg ggatgccggc gctcgaagta gcgagtctgc tgctccatgc aagccagcca
     4501 gggccgccag aagaagatgt tggccacctc gtactgactg tccccgaaca tagcttcgga
     4561 ccagtcgatt acggcggtga tgcggccgtt gtcggtcagg acgttgttgc tgccgaagtc
     4621 ggcatggacg aggtggcgca cctcgggaca gtcttcggcc cacagcatca gttcgtccag
     4681 ggcttgagct acgctggcgg acacggtgtc gtccatcacg gtctgccagt ggtagacatg
     4741 gggatcagca atggcgcaaa tgaaatcccg ccaggttgtg tactggccga tgccttgggg
     4801 cccgaatggg ccgaagccgc tggtttgact gaggtcggcg gcggcaatag catccatagc
     4861 ttcggcgaca ggctgtaaca cagcaggcag ctctgtttcg gggaggtctt ggagagtaac
     4921 gccttgggcg cgtctactga tgcagtatgt caggctctcg ctgaactcgc cgatgtccaa
     4981 cacttcgggg atgggtagtg cagcgctggc gaagtggcgg tacacgtagc ggtctttgta
     5041 gaagccatca gcgcagctat tcacccgcag tacatagccg cgtccgccga catcgaagct
     5101 gaaggctcgg ctctcttcgc cctccgacaa ctgcatcagg tcgctcacac tgtcgaactt
     5161 ctcgatgaga aatttttcaa cgctggtagc ggtgagttcg ggcttcttca tggtggcgct
     5221 agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     5281 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     5341 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     5401 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5461 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     5521 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     5581 ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg
     5641 ccatggtgct gcgc
//
